The rabbit isolated iris sphincter muscle maintained in an isotonic state is unaffected by applied serotonin (5-hydroxytryptamine or 5-HT) whereas carbachol causes the muscle to contract. Serotonin does, however, produce a relaxation of the contracted muscle in a dose-dependent manner. This effect is also induced by the 5-HT receptor agonists 8-OH-DPAT (8-hydroxy-2-[di-n-propyl-amino] tetralin, RU 24969 (5-methoxy-3-[l, 2, 3, 6, tetrahydro-4-pyridinyl]-1-indole) and ipsapirone, suggesting the involvement of 5-HT1Areceptors. This view is supported by the finding that metergoline, methysergide and propranolol all counteracted the effect produced by serotonin. While 5-HT3receptors are not involved in the described process, a minor involvement of 5-HT2receptors cannot be excluded as methysergide partially counteracted the serotonin response. These data provide evidence that serotonin receptors, in particular the 5-HT1Asubtype, may be associated with the iris sphincter muscle and suggest their involvement in the regulation of pupil size.